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Manufacturing, Control, and Performance Evaluation of a Gecko-Inspired Soft Robot
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Antibody engineering to improve manufacturability.

Sujeewa D Wijesuriya1, Elizabeth Pongo2, Milan Tomic1

  • 1Ology Bioservices, Inc, 626 Bancroft, Suite D, Berkeley, CA, 94710, USA.

Protein Expression and Purification
|April 16, 2018
PubMed
Summary
This summary is machine-generated.

Antibody light chain (LC) engineering resolved poor expression and aggregation issues in Chinese Hamster Ovary (CHO) cells. Pairing heavy chains (HC) with new LCs improved antibody production and quality while maintaining binding affinity.

Keywords:
Antibody developabilityAntibody expressionAntibody manufacturabilityHeavy and light chain pairingHeavy chain: light chain ratioSize exclusion chromatography

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Area of Science:

  • Biotechnology
  • Protein Engineering
  • Mammalian Cell Culture

Background:

  • Antibody expression in Chinese Hamster Ovary (CHO) cells can vary significantly.
  • A specific human monoclonal antibody, mAb B-c, exhibited poor expression and high aggregation levels, posing manufacturing challenges.

Purpose of the Study:

  • To identify the source of manufacturability issues for mAb B-c.
  • To engineer an alternative light chain (LC) to improve antibody expression and product quality.

Main Methods:

  • Transient and stable expression in CHO-K1 cells were used to assess antibody production.
  • Light chain shuffling was performed with the original heavy chain (HC) to identify novel LCs.
  • Binding affinity, expression levels, and aggregate formation were evaluated.

Main Results:

  • The light chain (LC) was identified as the primary cause of mAb B-c's manufacturability problems.
  • Antibodies engineered with new LCs demonstrated high expression at a 1:1 or 2:1 LC:HC ratio, comparable to other antibodies.
  • The new LCs significantly reduced high molecular weight aggregates and heavy chain (HC) dimer formation.

Conclusions:

  • Light chain engineering is a viable strategy to overcome antibody manufacturability issues.
  • Pairing heavy chains with alternative light chains can enhance antibody expression and product quality.
  • This approach maintains or improves antibody binding affinity.